Development of a Two-Layer Porous Scaffold Based on Porcine Nasal Septal Cartilage for Orthopedics.

The aim of the study was to design a construct based on a nasal septal cartilage plate providing required cell differentiation in different layers to replace a deep osteochondral defect and develop an algorithm of chemical and physical effect sequence to create non-immunogenic two-layer porous structure with requisite elasto-mechanical properties. MATERIALS AND METHODS: The plates derived from porcine nasal septal hyaline cartilage covered by perichondrium were multi-stage treated including freezing, equilibrating in a hypotonic saline solution (type I specimens); trypsinization, point IR-laser effect, re-trypsinization (type II specimens); a stabilizing effect of crosslinking agents - glyceraldehyde/ribose in an acidic medium - washing (type III specimens).For all type specimens:there were established stability parameters (collagen denaturation temperature using a thermal analysis; and Young's modulus using a mechanical analysis);there were determined morphological characteristics using light and polarization microscopy with classical staining and nonlinear optical microscopy in second-harmonic generation mode. RESULTS: Thermal, mechanical, and morphological properties in type I specimens slightly differed from those of the initial nasoseptal system. A considerable part of cells had destroyed membranes.In type II specimens, thermal stability of collagen frame was significantly lower; Young's modulus decreased more than fourfold compared to type I specimens. Collagen structure of hyaline cartilage appeared to be disarranged, although the morphological differences of the hyaline part and perichondrium preserved. The construct matrix was almost completely decellularized. Successive exposure to laser radiation and trypsin resulted in the formation of partial holes in the matrix, ~100 µm in diameter.In type III specimens, both the thermal stability of the collagen frame and Young's modulus (E) increased. Glyceraldehyde was more effective than ribose, E having reached the value typical for intact hyaline cartilage. Collagen fibers in type III specimens were thicker than in type I and II specimens. The morphological differences of the hyaline part and perichondrium and partial holes were preserved. CONCLUSION: Due to sequential treatment by salts, trypsin, IR-laser radiation, and nontoxic crosslinking agents, nasal septal cartilage plate forms porous acellular construction consisting of two layers formed by type I (from perichondrium) and type II (from hyaline part) collagen fibers. In the present construction, stability, mechanical properties, and size of the partial holes can be assigned for cell colonization. It enables to use the construction to replace articular cartilage defects.

Age-related adaptive responses of mitochondria of the retinal pigment epithelium to the everyday blue LED lighting.

The effect of everyday blue light (λ = 440-460 nm) on mitochondria of the retinal pigment epithelium of different age groups of Japanese quail was studied using electron microscopy, morphometric methods, and biochemical analysis. We have found a significant increase in the number of mitochondria, including those modified, mainly in young birds. In addition, cell metabolic activity increased in response to blue lighting. These changes are assumed to reflect an adaptive response of mitochondria aimed at neutralizing the phototoxic effect of blue light caused by accumulation of lipofuscin granules.

Photoinduced changes in subcellular structures of the retinal pigment epithelium from the Japanese quail Coturnix japonica.

Fifteen-week-old sexually mature female Japanese quails (Coturnix japonica) grown under various lighting conditions were used in the study. It was found that the number of mitochondria and phagosomes was increased by 1.5-fold in the retinal pigment epithelium from birds reared for 95 days under blue light (440-470 nm) vs. reduced blue light component conditions. Also, it was found that egg production was increased by 15% in birds reared under blue light compared to other lightning conditions. Thus, we concluded that blue light conditions resulted in elevating metabolic activity and accelerating pace of life in Japanese quails. It is assumed that the blue light-induced effects are probably due to inhibition of melatonin synthesis.

[The estimation of age-related sensitivity of the retinal pigment epithelium of Japanese quail (Coturnix japonica) to the light damage].

The effect of blue light damage (445-455 nm, 4 J/cm2) to retinal pigment epithelium (RPE) subcellular structures was investigated in 4 age risk groups (9, 25, 40 and 52 weeks) of Japanese quail Coturnix japonica by light and electron microscopy. The indicator of biological aging of RPE was age-related accumulation of lipofuscin granules: 5-6-fold increase in their quantity increasing by 5-6 times in quails at 52 weeks. The main photo-induced changes observed after 24 h of the photo radiation were located in the blood-retinal barrier, such as loss of homogeneity of Bruch's membrane, disorganization of basal processes, deformations of the nuclei and mitochondria shapes. Those effects ofphotobleaching were more expressed in young birds. But for the older 52-week age birds it was not so noticeable, because their retinal pigment epithelium structures had disorders which were similar to those in younger birds after photodamage.